Adolescent ; Chondrosarcoma ; pathology ; surgery ; Douglas' Pouch ; pathology ; surgery ; Female ; Humans ; Peritoneal Neoplasms ; pathology ; surgery ; Soft Tissue Neoplasms ; pathology ; surgery

To investigate the secondary metabolites of endophytic fungi Pericinia sp. F-31. Column chromatography on silica gel, Sephadex LH-20 and semi-preparative HPLC were used to separate and purify the compounds. Two compounds were isolated from the fermentation broth of Periconia sp. Their structures were identified as 5-(1-hydroxyhexyl) -6-methyl-2H-pyran-2-one (1) and 2-(3-hydroxy-4-methylphenyl) -propanoic acid (2). Compound 1 was a new lactone compound, compound 2 was new natural product, and the NMR data of compound 2 was reported for the first time.
Annona ; microbiology ; Ascomycota ; chemistry ; genetics ; isolation & purification ; metabolism ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; metabolism ; Endophytes ; chemistry ; genetics ; isolation & purification ; metabolism ; Lactones ; chemistry ; isolation & purification ; metabolism ; Mass Spectrometry ; Molecular Structure

Objective The relationship between circular RNAs (circRNA) and lung adenocarcinoma (LAC) remains to be further explored. This study aimed to investigate the expression of Hsa_circ_0002360 in LAC and its value in the clinical diagnosis and treatment of the malignancy. Methods This study included 50 cases of LAC treated in our hospital from May 2018 to June 2019 and another 50 healthy subjects as normal controls. We determined the expression of Hsa_circ_0002360 in the LAC cells by qRT⁃PCR. We cultured human LAC cell lines A549, H1299 and H1975 and human bronchial mucosal epithelial cell line BEAS-2B, inoculated the A549 and H1975 LAC cells in the 6-well plate, with 3 wells for si-circRNAs and the other 3 for negative controls (si-NC), followed by cell transfection experiment. Using the CCK8, colony formation and wound healing assays, we detected the effects of Hsa_circ_0002360 on the activity, proliferation, invasiveness and metastasis of the A549 and H1975 cells. Results The expression of Hsa_circ_0002360 was significantly higher in the LAC tissue than in the adjacent tissue (2.84 ± 0.12 vs 1.46 ± 0.08, P < 0.01), and so was it in the serum of the LAC patients than in the normal controls (2.59 ± 0.17 vs 1.21 ± 0.11, P < 0.01), but remarkably lower in the serum of the patients after than before surgery (1.58 ± 0.09 vs 2.65 ± 0.13, P < 0.01). Besides, its expression was significantly increased in the LAC A549, H1299 and H1975 cell lines compared with that in the BEAS-2B cells (P < 0.05). In comparison with the si-NC group, the si-circRNA group showed significant decreases in the rates of colony formation ([65.66± 4.93]% vs [38.33 ± 1.69]%, P < 0.01) and migration of the cells ([54.66±4.10]% vs [13.50 ± 3.34]%, P < 0.01) and in the count of H1975 cells as well (P < 0.01). Conclusion Hsa_circ_0002360 can be used as a biomarker for the early diagnosis and treatment of lung adenocarcinoma.

In the research and development of new drugs, it is very important to investigate the in vitro metabolism of candidate drugs. Traditional models such as liver microsomes have many limitations, while the in vitro model of recombinant human drug metabolizing enzymes is considered as an important and useful approach because of its convenient access, stable activity and low cost. In this study, six major human UDP-glucuronosyltransferases (UGTs) genes (UGT1A1, 1A3, 1A4, 1A6, 1A9 and 2B7) were cloned from human liver cDNA and heterologously expressed in Saccharomyces cerevisiae and baculovirus-infected insect cell. UGT1A1, 1A3, 1A6 and 1A9 were successfully expressed in yeast and showed glucuronidation activity against a variety of different structural types of substrates, but their activities were low. All six UGTs were successfully expressed and exhibited significantly improved glucuronidation activity when Trichopolusia ni cells BTI-TN5B1-4 (High Five) were used as the host. The recombinant human UGTs expressed in insect cells can catalyze the glucuronidation of their specific substrates, and the glucuronidation products were synthesized at milligram-scale with yields of 13%-66% for the first time, of which the structures were identified via MS, ¹H NMR, and ¹³C NMR spectroscopic analysis. Above all, the recombinant human UGTs yeast and insect cell expression systems constructed in this study can be used for in vitro metabolism evaluation in the early stage of new drugs research and development, and also provide a new tool for the synthesis of glucuronide metabolites.

OBJECTIVETo investigate the protein expression of caveolin-1 in type II alveolar epithelial cells (A549) exposed to carbon black nanoparticles (CB NPs) and the role of caveolin in the endocytosis of CB NPs.

METHODSA549 cells were exposed to 0, 25, 50, 100, 200, and 400 µg/ml CB NPs for 24 h; then, trypan blue assay was applied to determine the cell viability. A549 cells were also exposed to 0, 25, 50, and 100 µg/ml CB NPs for 24 h, then, transmission electron microscopy (TEM) and flow cytometry were applied to observe the morphological change of cells and cellular side scatter (SSC), and Western blot was used to analyze the effect of CB NPs on the protein expression of caveolin-1. A549 cells were co-exposed to1 µg/ml filipin and 100 µg/ml CB NPs for 24 h, then, the cellular SSC was observed.

RESULTSCompared with controls, the A549 cells exposed to 200 and 400 µg/ml CB NPs had the cell viability decreased by 38.2% and 46.6%, respectively (P < 0.05), while those exposed to 25, 50, and 100 µg/ml CB NPs showed no significant decrease in cell vitality (P > 0.05). The protein expression of caveolin-1 was significantly higher in the cells exposed to 50 and 100 µg/ml CB NPs than in controls (P < 0.05). The TEM showed that plasmalemmal vesicles containing black particles were found in the cytoplasm of the cells exposed to 50 and 100 µg/ml CB NPs. The flow cytometry showed that the cellular SSC ratio increased from 1.007 to 1.331 as the dose of CB NPs rose within 0 ∼ 100 µg/ml and fell to 1.25 after the cells were co-exposed to1 µg/ml filipin and 100 µg/ml CB NPs.

CONCLUSIONCarbon black nanoparticles can be transferred into A549 cells by endocytosis, but caveolin-mediated endocytic pathway plays a minor role in this process.

Caveolin 1 ; physiology ; Cell Line ; Endocytosis ; Humans ; Nanoparticles ; Soot ; pharmacokinetics

OBJECTIVETo investigate the biochemical metabolic changes detected by phosphorus-31 MR spectroscopy ((31)P MRS) with pathologic changes in the liver of fasting rabbits.

METHODSA total of 22 rabbits were under the starvation up to death to establish animal models. Hepatic (31)P MRS was performed in different period of 10 rabbits including normal condition, over-starvation, agonal condition and death after 30 min. Other 9 rabbits were divided into three type including over-starvation, agonal condition and death group with 3 rabbits in each group, and 3 healthy rabbits served as controls. All the 12 rabbits were sacrificed for the hepatic pathological examination. The MR examination was performed on a 1.5 T imager using a 1H/31P surface coil by the 2D chemical shift imaging technique. The relative quantities of phosphomonoesters (PME), phosphodiesters (PDE), inorganic phosphate (Pi) and adenosine triphosphate (ATP) were measured.

RESULTSAll the relative quantification of phosphorus metabolites were changed significantly from starvation to death (X(2)=23.13-35.41, P<0.01). The relative quantifications of ATP of normal condition, over-starvation, agonal condition and death were 2.54 +/-0.53, 1.73 +/-0.14, 0.88 +/-0.23 and 0.05 +/-0.08, respectively (rs=1.0, P<0.01). The relative quantifications of PDE from normal to death were 1.25 +/-0.54, 2.76 +/-0.23, 3.33 +/-0.49 and 3.87 +/-0.43, respectively, and those of Pi were 0.42 +/-0.02, 0.65 +/-0.05, 0.89 +/-0.15 and 0.99 +/-0.08, respectively (rs=1.0, P <0.01). The relative quantifications of PME were also significantly changed (rs=0.4, P=0.6). The pathologic changes of normal condition, over-starvation, agonal condition and death: decreased size of hepatocytes, loss of cell number, cellular swelling, degeneration and cell necrosis or hepatic hemorrhage became more and more pronounced.

CONCLUSION(31)P MRS can monitor dynamic changes of relative quantification of phosphorus metabolites, which are correlated with the pathological severity of acute hepatic injury by fasting.

Animals ; Death ; Dose-Response Relationship, Radiation ; Female ; Liver ; metabolism ; pathology ; Magnetic Resonance Spectroscopy ; methods ; Male ; Phosphorus ; metabolism ; Phosphorus Isotopes ; metabolism ; Rabbits ; Random Allocation ; Starvation

OBJECTIVE: To investigate the effect of diet-induced hyperlipidemia on TGF-beta/Smad signaling pathway in the kidney of diabetic rats, and to explore the mechanism by which hyperlipidemia leads to renal injury in diabetes. METHODS: Diabetic rats and non-diabetic rats were fed with normal fat diet and high fat diet for 16 weeks, respectively. The expressions of TGF-beta1, TbetaRII, and Col-IV mRNA in the renal cortex were examined by reverse transcriptase-PCR,TbetaRII and p-Smad staining in glomerular cells were detected by immunohistochemical staining, and the expression of TGF-beta1 and Col-IV protein was determined by Western blot. RESULTS: Diet-induced hyperlipidemia up-regulated the levels of TGF-beta1, TbetaRII, p-Smad, and Col-IV protein and mRNA in the renal cortex of diabetic rats compared with those of non-diabetic rats. However, feeding high fat diet to non-diabetic rats had no influence on the expression of TGF-beta1, TbetaRII, p-Smad2, and Col-IV in the renal cortex. CONCLUSION Hyperlipidemia induced by high fat diet ingestion leads to renal injury in diabetic rats through activating TGF-beta1 /Smad signaling pathway.
Animals ; Diabetes Mellitus, Experimental ; complications ; metabolism ; Diabetic Nephropathies ; metabolism ; Dietary Fats ; administration & dosage ; Female ; Hyperlipidemias ; complications ; metabolism ; Kidney ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Smad Proteins ; metabolism ; Transforming Growth Factor beta1 ; metabolism

We report here on a rare case of an ectopic pancreatic tissue in the anterior mediastinum. A 32-year-old woman without any symptoms was transferred to our hospital because of an abnormal large mediastinal shadow on her chest radiograph during a checkup. The computed tomography (CT) scan revealed a giant cystic-solid mass that measured 16 x 13 x 8 cm and it was located in the center of the anterior mediastinum and it symmetrically grew to two sides. On enhanced CT scans, the solid component of the mass showed marked enhancement. We performed total surgical resection of the mass and complete pancreatic tissues were verified on the pathological examination.
Adult ; Choristoma/*radiography/surgery ; Diagnosis, Differential ; Female ; Humans ; Mediastinal Diseases/*radiography/surgery ; *Pancreas ; Tomography, X-Ray Computed

OBJECTIVETo study the effects of cadmium on hepatocellular DNA damage, expression of proto-oncogenes c-myc, c-fos, and c-jun as well as apoptosis in rats.

METHODSCadmium chloride at the doses of 5, 10, and 20 micromol/kg was given to rats by i.p. and there were 5 male SD rats in each group. Hepatocellular DNA damage was measured by single cell gel electrophoresis (or comet assay), while expression of proto-oncogenes c-myc, c-fos, and c-jun in rat hepatocytes were measured by Northern dot hybridization. C-Myc, c-Fos, and c-Jun were detected with immuno-histochemical method. Hepatocellular apoptosis was determined by TUNEL (TdT-mediated dUTP Nick End Labelling) and flow cytometry.

RESULTSAt the doses of 5, 10, and 20 micromol/kg, cadmium chloride induced DNA damage in rat hepatocytes and the rates of comet cells were 50.20%, 88.40%, and 93.80%, respectively. Results also showed an obvious dose-response relationship between the rates of comet cells and the dose of cadmium chloride (r = 0.9172, P < 0.01). Cadmium chloride at the doses of 5, 10, and 20 micromol/kg induced expression of proto-oncogenes c-myc, c-fos, and c-jun. The positive brown-yellow signal for c-myc, c-fos, and c-jun was mainly located in the cytoplasm of hepatocytes with immunohistochemical method. TUNEL-positive cells were detected in cadmium-treated rat livers. Apoptotic rates (%) of cadmium-treated liver cells at the doses of 5, 10, and 20 micromol/kg were (17.24 +/- 2.98), (20.58 +/- 1.35), and (24.06 +/- 1.77) respectively, being significantly higher than those in the control. The results also displayed an obvious dose-response relationship between apoptotic rates and the dose of cadmium chloride (r = 0.8619, P < 0.05).

CONCLUSIONCadmium at 5-20 micromol/kg can induce hepatocellular DNA damage, expression of proto-oncogenes c-myc, c-fos, and c-jun as well as apoptosis in rats.

Animals ; Apoptosis ; drug effects ; Cadmium ; toxicity ; DNA Damage ; Gene Expression Regulation ; drug effects ; Hepatocytes ; cytology ; drug effects ; metabolism ; Male ; Proto-Oncogene Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins c-fos ; genetics ; metabolism ; Proto-Oncogene Proteins c-jun ; genetics ; metabolism ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the effects of selenium and zinc on oxidative stress, apoptosis, and cell cycle changes in rat renal cells induced by fluoride.

METHODSWistar rats were given distilled water containing sodium fluoride (50 mg/L NaF) and were gavaged with different doses of selenium-zinc preparation for six months. Four groups were used and each group had eight animals (four males and four females). Group one, sham-handled control; group two, 50 mg/L NaF; group three, 50 mg/L NaF with a low dose of selenium-zinc preparation (0.1 mg/kg Na2 SeO3 and 14.8 mg/kg ZnSO4 x 7H2O); and group four, 50 mg/L NaF with a high dose of selenium-zinc preparation (0.2 mg/kg Na2 SeO3 and 29.6 mg/kg ZnSO4 x 7H2O). The activities of serum glutathione peroxidase (GSH-Px), kidney superoxide dismutase (SOD), and the levels of malondialdehyde (MDA) and glutathione (GSH) in the kidney were measured to assess the oxidative stress. Kidney cell apoptosis and cell cycle were detected by flow cytometry.

RESULTSNaF at the dose of 50 mg/L increased excretion of fluoride in urine, promoted activity of urine gamma-glutamyl transpeptidase (gamma-GT), inhibited activity of serum GSH-PX and kidney SOD, reduce kidney GSH content, and increased kidney MDA. NaF at the dose of 50 mg/L also induced rat renal apoptosis, reduced the cell number of G2/M phase in cell cycle, and decreased DNA relative content significantly. Selenium and zinc inhibited effects of NaF on oxidative stress and apoptosis, promoted the cell number of G2/M phase in cell cycle, but failed to increase relative DNA content significantly.

CONCLUSIONSodium fluoride administered at the dose of 50 mg/L for six months induced oxidative stress and apoptosis, and changes the cell cycle in rat renal cells. Selenium and zinc antagonize oxidative stress, apoptosis, and cell cycle changes induced by excess fluoride.

Animals ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Glutathione ; metabolism ; Glutathione Peroxidase ; blood ; Kidney ; drug effects ; metabolism ; Malondialdehyde ; metabolism ; Oxidative Stress ; drug effects ; Rats ; Rats, Wistar ; Selenium ; pharmacology ; Sodium Fluoride ; antagonists & inhibitors ; toxicity ; urine ; Superoxide Dismutase ; metabolism ; Zinc ; pharmacology ; gamma-Glutamyltransferase ; urine